Preparation of β2AR-T4L and Nb80 are described in Supplementary Methods . BI-167107 bound β2AR-T4L and Nb80 preincubated in 1:1.2 molar ratio were mixed in monoolein containing 10% cholesterol in 1:1.5 protein to lipid ratio (w/w). Initial crystallization leads were identified and optimized in 24-well glass sandwich plates using 50 nL protein:lipid drops overlaid with 0.8 μl precipitant solution in each well and sealed with a glass cover slip. Crystals for data collection were grown at 20° C in hanging-drop format using 0.8 μl reservoir solution (36 to 44% PEG 400, 100 mM Tris pH 8.0, 4 % DMSO, 1 % 1,2,3-heptanetriol) diluted 2 to 4-fold in water. Crystals grew to full size, typically 40 x 5 x 5 μm3, within 7 to 10 days. Crystals were flash frozen and stored in liquid nitrogen with reservoir solution as cryoprotectant. Diffraction data collection and processing, and structure solution and refinement are described in Supplementary Methods .
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Monoolein
Monoolein
Monoolein is a naturally occurring lipid compound that plays a crucial role in various biological and chemical processes.
It is commonly used as a surfactant, emulsifier, and in the formulation of drug delivery systems.
PubCompare.ai's AI-driven protocol comparison tool can help researchers optimize their Monoolein research by identifying the most accurate and reproducible procedures from literature, preprints, and patents.
This allows for enhanced research efficiency and accuracy, boosting the overall quality of Monoolein-related investigations.
Experienec the future of research optimization today with PubCompare.ai.
It is commonly used as a surfactant, emulsifier, and in the formulation of drug delivery systems.
PubCompare.ai's AI-driven protocol comparison tool can help researchers optimize their Monoolein research by identifying the most accurate and reproducible procedures from literature, preprints, and patents.
This allows for enhanced research efficiency and accuracy, boosting the overall quality of Monoolein-related investigations.
Experienec the future of research optimization today with PubCompare.ai.
Most cited protocols related to «Monoolein»
BI167107
Cholesterol
Cryoprotective Agents
Crystallization
Freezing
Lipids
Molar
monoolein
Nitrogen
polyethylene glycol 400
Proteins
Sulfoxide, Dimethyl
Tromethamine
hKOR-T4L was expressed in Spodoptera frugiperda (Sf9) cells. Ligand-binding and functional assays were performed as described in Methods. Sf9 cells were solubilized using 1% (w/v) n-dodecyl-β-D-maltopyranoside (DDM) and 0.2% (w/v) cholesteryl hemisuccinate (CHS), and purified by immobilized metal ion affinity chromatography (IMAC), followed by reverse IMAC after cleaving N-terminal FLAG-10xHis tags by His-tagged Tobacco Etch Virus (TEV) protease. The purified protein was mixed with monoolein and cholesterol in a ratio of 40%:54%:6% (w/w) to form lipidic cubic phase (LCP) from which the receptor was crystallized. Crystals were grown at 20 °C in 45 nl protein-laden LCP boluses overlaid by 800 nl of precipitant solutions as described in Methods. Crystals were harvested from the LCP matrix and flash frozen in liquid nitrogen. X-ray diffraction data were collected on the 23ID-B/D beamline (GM/CA CAT) at the Advanced Photon Source, Argonne, IL using a 10 μm minibeam at wavelength of 1.0330 Å. Data collection, processing, structure solution and refinement are described in Methods. Modeling of JDTic analogues and hKOR-selective morphine derivatives nor-BNI and GNTI was performed using ICM-Pro; SYBYL-X 1.3 and GOLD Suite 5.1 were used to model RB-64 complexes, as described in Methods.
Full Methods and any associated references are available in the online version of the paper at www.nature.com/nature .
Biological Assay
Cholesterol
cholesterol-hemisuccinate
Chromatography, Affinity
Cuboid Bone
Freezing
Gold
Ligands
Lipids
Metals
monoolein
Morphines
Nitrogen
Protein C
Proteins
RB-64
Sf9 Cells
Spodoptera frugiperda
TEV protease
X-Ray Diffraction
ammonium phosphate
Biological Assay
Cholesterol
cholesterol-hemisuccinate
Chromatography, Affinity
Citrate
Crystallization
Cuboid Bone
Diamond
Doxepin
Freezing
Komagataella pastoris
Ligands
Lipids
Magnesium Chloride
Metals
monoolein
Nitrogen
polyethylene glycol 400
Proteins
Saccharomyces cerevisiae
TEV protease
Tissue, Membrane
TNFSF14 protein, human
X-Ray Diffraction
FAUC50-β2ARH93CT4L was expressed, purified and crystallized as described in Methods. The receptor was crystallized in a cholesterol-doped (10%) monoolein cubic phase overlaid with precipitant in glass sandwich plates. Optimized precipitant consisted of 24–27 %(v/v) PEG400, 200 mM Li2SO4, 4 %(v/v) DMSO, 3.5 %(v/v) 1,4-butanediol, 100 mM MES pH 6.7. After 3–5 days of growth, crystals were harvested after adding an excess of precipitant solution for cryoprotection, and flash frozen in liquid nitrogen.
Butylene Glycols
Cholesterol
Cuboid Bone
Freezing
monoolein
Nitrogen
polyethylene glycol 400
Sulfoxide, Dimethyl
BRIL-NOP was expressed in Spodoptera frugiperda (Sf9) insect cells. Ligand binding asays were performed as described in Methods online. Sf9 membranes were solubilized using 0.5% n-dodecyl-β-D-maltopyranoside (w/v) and 0.1% cholesteryl hemisuccinate (w/v), and purified by immobilized metal ion affinity chromatography (IMAC). Receptor crystallization was performed by the lipidic cubic phase (LCP) method. The protein-LCP mixture contained 40% (w/w) concentrated receptor solution, 54% (w/w) monoolein, and 6% (w/w) cholesterol. Crystals were grown in 40 nL protein-laden LCP bolus overlaid by 0.8 μL of precipitant solution (25–30% (v/v) PEG 400, 100–200 mM potassium sodium tartrate tetrahydrate, 100 mM BIS-TRIS propane [pH 6.4]) at 20 °C. Crystals were harvested directly from LCP matrix and flash frozen in liquid nitrogen. X-ray diffraction data were collected at 100 K on the 23ID-B/D beamline (GM/CA CAT) of the Advanced Photon Source at the Argonne National Laboratory using a 10 μm collimated minibeam. Diffraction data from 23 crystals were merged for the final dataset. Data collection, processing, structure solution and refinement are described in Methods online.
Full Methods and any associated references are available in the online version of the paper at (web).
1,3-bis(tris(hydroxymethyl)methylamino)propane
Cells
Cholesterol
cholesterol-hemisuccinate
Chromatography, Affinity
Crystallization
Cuboid Bone
Freezing
Insecta
Ligands
Lipids
Metals
monoolein
Nitrogen
polyethylene glycol 400
Proteins
sodium potassium tartrate
Spodoptera frugiperda
Tissue, Membrane
X-Ray Diffraction
Most recents protocols related to «Monoolein»
Cold pressed grapeseed oil was purchased from local grocery store (Athens, GA, USA) . Capric acid (≥ 99％ purity) , monoolein, 2-monoolein, diolein, GLC-461 fatty acid methyl ester (FAME) were purchased from Nu-check Prep, Inc. (Elysian, MN, USA) . Lipozyme ® RM IM (Rhizomucor miehei lipase immobilized on a resin carrier with specific activity of 442 IUN g-1 (interesterification unit per gram) as specified by manufacturer) and Lipozyme ® 435 (recombinant lipase from Candida antarctica, expressed on Aspergillus niger, and immobilized on a macroporous hydrophobic resin with specific activity of 10,000 PLU g-1 ［propyl laurate unit per gram］ as specified by the manufacturer) were purchased from Novozymes North America, Inc. (Franklinton, NC, USA) . Mixed tocopherols standard was purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA) , and contained 11.13, 1.55, 68.59, and 18.73％ of α, β, γ, and δ-tocopherols (w/w) .
All other analytical and HPLC grade reagents, chemicals, standards, and solvents used were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA) , Tokyo Chemical Industry Co. (TCI) America Division (Portland, OR, USA) , and Fisher Chemical (Fair Lawn, NJ, USA) .
All other analytical and HPLC grade reagents, chemicals, standards, and solvents used were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA) , Tokyo Chemical Industry Co. (TCI) America Division (Portland, OR, USA) , and Fisher Chemical (Fair Lawn, NJ, USA) .
Monoolein was purchased from Croda (Cithrol
GMO HP-SO-LK, purity >96%). Sodium hydroxide (Honeywell), hydrochloric
acid (Honeywell), urea (ultrapure 99%, Alfa Aesar), and urease (U4002–100
KU, Jack Beans, 100,000 units/g solid) were used as received. Methyl
formate (anhydrous, 99%), sodium phosphate monobasic, and sodium phosphate
dibasic were purchased from Sigma. All solutions were prepared using
Milli-Q water (18.2 MΩ cm–1, Millipore, Bedford,
MA). pH-adjusted 100 mM sodium phosphate buffer solutions were used
throughout the experiment.
GMO HP-SO-LK, purity >96%). Sodium hydroxide (Honeywell), hydrochloric
acid (Honeywell), urea (ultrapure 99%, Alfa Aesar), and urease (U4002–100
KU, Jack Beans, 100,000 units/g solid) were used as received. Methyl
formate (anhydrous, 99%), sodium phosphate monobasic, and sodium phosphate
dibasic were purchased from Sigma. All solutions were prepared using
Milli-Q water (18.2 MΩ cm–1, Millipore, Bedford,
MA). pH-adjusted 100 mM sodium phosphate buffer solutions were used
throughout the experiment.
Full text: Click here
Fluorometholone was procured from Festiva Pharma in Gujarat. Glyceryl Monoolein and Poloxamer 407 were graciously provided as gift samples by Mohini Organics and BASF, Mumbai, respectively. Porcine stomach mucin, ethanol, sodium bicarbonate, sodium chloride and other reagents were acquired from Sigma-Aldrich® Inc. in the USA. Dialysis membranes with molecular weight cut-offs of 12,000-14,000 were purchased from Sigma Aldrich in Darmstadt, Germany. All other chemicals and solvents were of analytical grade and used without additional purification. Deionized, distilled water was utilized for all purposes and animal organs were obtained from the slaughterhouse.
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Open the protocol to access the free full text link
MO was heated
and sonicated at 60 °C in a water bath for 20 min to allow the
transformation from solid to liquid/fluid-like.15 (link) After this, 94 mg (approximately 100 μL) of MO was
pipetted and subsequently mixed with 119 μL of ethanol to yield
a mixture of 50/50 (w/w) monoolein/ethanol. This mixture was vortexed
for 5 min to homogenize the sample. The procedure above was repeated
for oleic acid (OA) to produce OA/EtOH, 50/50 (w/w). All samples were
equilibrated at room temperature before use.
To prepare 10%OA/MO
(w/w), 20 μL of 50%OA/EtOH was added to 200 μL of 50%MO/EtOH,
followed by vortexing for 5 min. The lipid ethanol matrix was dried
in a fume hood for 48 h to allow solvent evaporation. The Eppendorf
tube that held the sample was weighed before and after evaporation
to determine solvent loss.
and sonicated at 60 °C in a water bath for 20 min to allow the
transformation from solid to liquid/fluid-like.15 (link) After this, 94 mg (approximately 100 μL) of MO was
pipetted and subsequently mixed with 119 μL of ethanol to yield
a mixture of 50/50 (w/w) monoolein/ethanol. This mixture was vortexed
for 5 min to homogenize the sample. The procedure above was repeated
for oleic acid (OA) to produce OA/EtOH, 50/50 (w/w). All samples were
equilibrated at room temperature before use.
To prepare 10%OA/MO
(w/w), 20 μL of 50%OA/EtOH was added to 200 μL of 50%MO/EtOH,
followed by vortexing for 5 min. The lipid ethanol matrix was dried
in a fume hood for 48 h to allow solvent evaporation. The Eppendorf
tube that held the sample was weighed before and after evaporation
to determine solvent loss.
Full text: Click here
Top products related to «Monoolein»
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Monoolein is a lipid compound used in the development and production of various types of laboratory equipment. It serves as a key component in the formulation of lipid-based systems, such as liposomes and lipid nanoparticles, which are commonly utilized in research and pharmaceutical applications.
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MicroMounts are small, circular loops designed for the manipulation and mounting of protein crystals for X-ray crystallography experiments. They provide a stable and precise platform for holding and positioning the crystals during data collection.
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Oleic acid is a long-chain monounsaturated fatty acid commonly used in various laboratory applications. It is a colorless to light-yellow liquid with a characteristic odor. Oleic acid is widely utilized as a component in various laboratory reagents and formulations, often serving as a surfactant or emulsifier.
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Cholesterol is a lab equipment product that measures the concentration of cholesterol in a given sample. It provides quantitative analysis of total cholesterol, HDL cholesterol, and LDL cholesterol levels.
Sourced in United States, Germany, France, India, China, United Kingdom
Triolein is a laboratory reagent used as a standard for lipid analysis. It is a triglyceride composed of three oleic acid molecules esterified to a glycerol backbone. Triolein is commonly used as a reference substance in analytical techniques such as gas chromatography and high-performance liquid chromatography to quantify the composition of lipid samples.
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The RockImager 1000 is a laboratory equipment designed for crystal imaging. It is used to capture high-resolution images of crystalline samples.
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The Rock Imager is a laboratory instrument designed for automated imaging and analysis of crystallization experiments. It captures high-resolution images of crystal samples at user-defined intervals, providing a comprehensive visual record of the crystallization process.
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The Mosquito LCP robot is a liquid handling system designed for precise and automated pipetting tasks. It features a high-precision dispenser and liquid level detection capabilities to ensure accurate liquid transfer. The Mosquito LCP robot is suitable for a variety of laboratory applications that require precise liquid handling.
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1,3-diolein is a laboratory reagent used in various biochemical and analytical applications. It is a synthetic lipid that consists of two fatty acid chains esterified to a glycerol backbone. The primary function of 1,3-diolein is to serve as a model compound for the study of lipid metabolism and membrane structure and dynamics. It can be used in liposome and membrane-related research.
Monoolein is a lipid compound commonly used in laboratory settings for various applications. It is a monoacylglycerol with a single fatty acid chain attached to a glycerol backbone. Monoolein serves as a key component in the formulation of various lipid-based systems, such as liposomes, emulsions, and microemulsions, which are widely utilized in research and development across multiple scientific domains.
More about "Monoolein"
Monoolein, also known as glycerol monooleate or 1-monoolein, is a naturally occurring lipid compound that plays a crucial role in various biological and chemical processes.
It is a type of monoglyceride, which is a glycerol molecule with a single fatty acid chain attached.
Monoolein is commonly used as a surfactant, emulsifier, and in the formulation of drug delivery systems.
It is valued for its ability to form various liquid crystalline phases, including the cubic phase, which can be leveraged in the development of advanced drug delivery platforms.
Beyond its applications in pharmaceuticals, monoolein is also utilized in the food industry, cosmetics, and even in the production of biomaterials.
It is often studied in conjunction with related lipids and compounds, such as MicroMounts, oleic acid, cholesterol, and triolein.
Researchers investigating monoolein-related processes can benefit from tools like PubCompare.ai's AI-driven protocol comparison tool.
This innovative solution helps identify the most accurate and reproducible procedures from the scientific literature, preprints, and patents, optimizing research efficiency and accuracy.
When working with monoolein, researchers may also utilize specialized equipment like the RockImager 1000 or the Mosquito LCP robot, which are designed to facilitate the study of lipid-based systems and crystal formation.
By understanding the multifaceted nature of monoolein and the resources available to support its research, scientists can unlock new insights and drive advancements in fields ranging from drug delivery to biomaterial development.
Experienec the future of research optimization today with PubCompare.ai.
It is a type of monoglyceride, which is a glycerol molecule with a single fatty acid chain attached.
Monoolein is commonly used as a surfactant, emulsifier, and in the formulation of drug delivery systems.
It is valued for its ability to form various liquid crystalline phases, including the cubic phase, which can be leveraged in the development of advanced drug delivery platforms.
Beyond its applications in pharmaceuticals, monoolein is also utilized in the food industry, cosmetics, and even in the production of biomaterials.
It is often studied in conjunction with related lipids and compounds, such as MicroMounts, oleic acid, cholesterol, and triolein.
Researchers investigating monoolein-related processes can benefit from tools like PubCompare.ai's AI-driven protocol comparison tool.
This innovative solution helps identify the most accurate and reproducible procedures from the scientific literature, preprints, and patents, optimizing research efficiency and accuracy.
When working with monoolein, researchers may also utilize specialized equipment like the RockImager 1000 or the Mosquito LCP robot, which are designed to facilitate the study of lipid-based systems and crystal formation.
By understanding the multifaceted nature of monoolein and the resources available to support its research, scientists can unlock new insights and drive advancements in fields ranging from drug delivery to biomaterial development.
Experienec the future of research optimization today with PubCompare.ai.